Hepatitis C virus (HCV) is the major etiological agent of post-transfusion and community-acquired non-A non-B hepatitis worldwide. It is estimated that over 200 million people worldwide are infected by the virus. A high percentage of carriers become chronically infected and many progress to chronic liver disease, so-called chronic hepatitis C. This group is in turn at high risk for serious liver disease such as liver cirrhosis, hepatocellular carcinoma and terminal liver disease leading to death.
The mechanism by which HCV establishes viral persistence and causes a high rate of chronic liver disease has not been thoroughly elucidated. It is not known how HCV interacts with and evades the host immune system. In addition, the roles of cellular and humoral immune responses in protection against HCV infection and disease have yet to be established. Immunoglobulins have been reported for prophylaxis of transfusion-associated viral hepatitis, however, the Center for Disease Control does not presently recommend immunoglobulins treatment for this purpose. The lack of an effective protective immune response is hampering the development of a vaccine or adequate post-exposure prophylaxis measures, so in the near-term, hopes are firmly pinned on antiviral interventions.
Various clinical studies have been conducted with the goal of identifying pharmaceutical agents capable of effectively treating HCV infection in patients afflicted with chronic hepatitis C. These studies have involved the use of interferon-alpha, alone and in combination with other antiviral agents. Such studies have shown that a substantial number of the participants do not respond to these therapies, and of those that do respond favorably, a large proportion were found to relapse after termination of treatment.
Until recently, interferon (IFN) was the only available therapy of proven benefit approved in the clinic for patients with chronic hepatitis C. However the sustained response rate is low, and interferon treatment also induces severe side-effects (i.e. retinopathy, thyroiditis, acute pancreatitis, depression) that diminish the quality of life of treated patients. Recently, interferon in combination with ribavirin has been approved for patients non-responsive to IFN alone. However, the side effects caused by IFN are not alleviated with this combination therapy. Pegylated forms of interferons such as PEG-Intron® and Pegasys® can apparently partially address these deleterious side-effects but antiviral drugs still remain the avenue of choice for oral treatment of HCV.
Therefore, a need exists for the development of effective antiviral agents for treatment of HCV infection that overcome the limitations of existing pharmaceutical therapies.
HCV is an enveloped positive strand RNA virus in the Flaviviridae family. The single strand HCV RNA genome is approximately 9500 nucleotides in length and has a single open reading frame (ORF) encoding a single large polyprotein of about 3000 amino acids. In infected cells, this polyprotein is cleaved at multiple sites by cellular and viral proteases to produce the structural and non-structural (NS) proteins. In the case of HCV, the generation of mature nonstructural proteins (NS2, NS3, NS4A, NS4B, NS5A, and NS5B) is effected by two viral proteases. The first one, as yet poorly characterized, cleaves at the NS2-NS3 junction; the second one is a serine protease contained within the N-terminal region of NS3 (henceforth referred to as NS3 protease) and mediates all the subsequent cleavages downstream of NS3, both in cis, at the NS3-NS4A cleavage site, and in trans, for the remaining NS4A-NS4B, NS4B-NS5A, NS5A-NS5B sites. The NS4A protein appears to serve multiple functions, acting as a cofactor for the NS3 protease and possibly assisting in the membrane localization of NS3 and other viral replicase components. The complex formation of the NS3 protease with NS4A seems necessary to the processing events, enhancing the proteolytic efficiency at all of the sites. The NS3 protein also exhibits nucleoside triphosphatase and RNA helicase activities. NS5B is a RNA-dependent RNA polymerase that is involved in the replication of HCV.
A general strategy for the development of antiviral agents is to inactivate virally encoded enzymes that are essential for the replication of the virus.
The following is a list of patent application published in the last few years that disclose HCV NS3 protease inhibitor peptide analogs that are structurally different from the compounds of the present invention:
WO 98/17679 (published Apr. 30, 1998); WO 99/50230 (published Oct. 7, 1999); WO 01/74768 (published Oct. 11, 2001); WO 98/22496 (published May 28, 1998); U.S. Pat. No. 6,187,905; WO 97/43310 (published Nov. 20, 1997); WO 01/58929 (published Aug. 16, 2001); WO 01/77113 (published Oct. 18, 2001); WO 01/81325 (published Nov. 1, 2001); WO 98/46597 (published Oct. 22, 1998); WO 98/46630 (published Oct. 22, 1998); JP 10298151 (published Nov. 10, 1998); JP 11127861 (published May 18, 1999); JP 2001103993 (published Apr. 17, 2001); JP 11292840 (published Oct. 26, 1999); WO 99/38888 (published Aug. 5, 1999); WO 99/64442 (published Dec. 16, 1999); WO 00/31129 (published Jun. 2, 2000); WO 01/32691 (published May 10, 2001); U.S. Pat. No. 6,159,938 (published Dec. 12, 2000): WO 01/02424 (published Jan. 11, 2001); WO 01/07407 (published Feb. 1, 2001); WO 01/40262 (published Jun. 7, 2001); and WO 01/64678 (published Sep. 7, 2001).
Peptide analogs which inhibit the HCV NS3 protease have been disclosed in WO 00/09543 (published Feb. 24, 2000), WO 00/09558 (published Feb. 24, 2002), WO 00/59929 (published Oct. 12, 2000) and WO 02/060926 (published Aug. 8, 2002). The compounds of the present invention distinguish themselves by having a different chemical structure and by the surprising finding that they specifically inhibit HCV NS3 protease while showing insignificant inhibitory activity against other serine proteases. The WO 00/59929 discloses the corresponding terminal acid of the present compounds, which also exhibit specificity. WO 03/053349 published on Jul. 3, 2003 also discloses macrocyclic tripeptide inhibitors of hepatitis C virus. Nevertheless, the specific activity of the present compounds was unexpected.
One advantage of the present invention is that it provides tripeptide compounds that are inhibitory to the NS3 protease, an enzyme essential for the replication of the hepatitis C virus.
A further advantage of one aspect of the present invention resides in the fact that the compounds specifically inhibit the NS3 protease and do not show significant inhibitory activity against other serine proteases such as human leukocyte elastase (HLE), porcine pancreatic elastase (PPE), or bovine pancreatic chymotrypsin, or cysteine proteases such as human liver cathepsin B (Cat B).
Furthermore, the compounds are active in cell culture.